polyclonal rabbit antibody anti-cht1  (Genemed Synthesis)

Journal: Journal of Pain Research

Article Title: Upregulation of the high-affinity choline transporter in colon relieves stress-induced hyperalgesia

doi: 10.2147/JPR.S164186

Figure Lengend Snippet: Protein and mRNA levels of CHT1 in the colon. Notes: ( A ) Western blot analysis reveals that CHT1 protein expression is enhanced in WAS rats compared to control rats, whereas the elevated CHT1 expression is partly abolished after treatment with PDTC. ( B ) CHT1 mRNA levels in WAS-exposed rats are higher than those in control rats, and are considerably declined after treatment with PDTC. n of each group =6; * P <0.05, compared to control; # P <0.05, compared to the WAS group. Data are presented as mean ± SD and analyzed with the S-N-K post hoc test. Abbreviations: PDTC, ammonium pyrrolidinedithiocarbamate; WAS, water avoidance stress.

Article Snippet: The sections were incubated with rabbit anti-rat polyclonal antibody raised against CHT1 (Millipore, Darmstadt, Germany) at room temperature for 24 hours, followed by incubation with the appropriate secondary antibodies.

Techniques: Western Blot, Expressing

Journal: Journal of Pain Research

Article Title: Upregulation of the high-affinity choline transporter in colon relieves stress-induced hyperalgesia

doi: 10.2147/JPR.S164186

Figure Lengend Snippet: Immunostaining of CHT1 in the colon. Notes: ( A ) Immunohistochemical staining for CHT1 reveals that the AOD of CHT1 in the muscular layer of WAS rats (right) is increased compared to that of control rats (left) (200×). ( B ) Immunofluorescence staining for CHT1 shows that the density of CHT1 in the muscular layer is significantly elevated after 10 days of WAS (200×). Both the black and white arrows indicate CHT1-positive cells that are mainly fusiform in shape and found within the circular muscle, exhibiting the morphologic characteristics of ICC and smooth muscle cells. Both black and white arrowheads indicate CHT1-positive cells that are predominantly round in shape, located between the circular muscle and longitudinal muscle, showing the hallmark of MP neurons. Bars: (A) = 100 µm, (B) = 100 µm; n of each group = 6. Abbreviations: AOD, average optical density; ICC, interstitial cells of Cajal; MP, myenteric plexus; WAS, water avoidance stress.

Article Snippet: The sections were incubated with rabbit anti-rat polyclonal antibody raised against CHT1 (Millipore, Darmstadt, Germany) at room temperature for 24 hours, followed by incubation with the appropriate secondary antibodies.

Techniques: Immunostaining, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Journal of Pain Research

Article Title: Upregulation of the high-affinity choline transporter in colon relieves stress-induced hyperalgesia

doi: 10.2147/JPR.S164186

Figure Lengend Snippet: Effect of MKC-231 on pain threshold and CHT1 expression in WAS rats. Notes: ( A ) Immunoblots (left) and a histogram (right) for CHT1 in the colon of WAS and WAS-MKC-231-treated rats reveal that the ratios of CHT1 to β-actin expression in WAS rats are considerably increased after 10 consecutive days of MKC-231 treatment. ( B ) The ACh content of WAS rats is higher than that of the control rats, but lower than that of the MKC-231-treated rats. ( C ) The AWR score of WAS + MKC-231 rats is significantly reduced in contrast to the WAS group. ( D ) Withdrawal events for the WAS + MKC-231 group in the VFF test are declined compared to those in the WAS group. ( E ) VMR amplitude of the WAS + MKC-231 group is lower than that of the WAS rats. n of each group = 6; * P <0.05, compared to the WAS group. Data are presented as mean ± SD and analyzed by two-way repeated-measures ANOVA with distention pressure and filament force as the repeated measures. Abbreviations: AWR, abdominal withdrawal reflex; CRD, colorectal distension; VFF, Von Frey filament; VMR, visceromotor response; WAS, water avoidance stress.

Article Snippet: The sections were incubated with rabbit anti-rat polyclonal antibody raised against CHT1 (Millipore, Darmstadt, Germany) at room temperature for 24 hours, followed by incubation with the appropriate secondary antibodies.

Techniques: Expressing, Western Blot

Journal: Journal of Pain Research

Article Title: Upregulation of the high-affinity choline transporter in colon relieves stress-induced hyperalgesia

doi: 10.2147/JPR.S164186

Figure Lengend Snippet: Distribution of CHT1 in the MP of colon. Notes: Immunofluorescence for CHT1, PGP9.5, and merged picture on whole-mount preparations of colonic MP from control ( A ), WAS ( B ), and WAS + MKC-231 rats ( C ) (200×) was displayed. The percentage of CHT1-positive neurons in the MP of WAS is significantly higher than that in the control group. In addition, the percentage of CHT1-positive neurons in the WAS + MKC-231 group is enhanced compared to that in the WAS group. Bars = 100 µm; n of each group = 6. Abbreviations: MP, myenteric plexus; PGP9.5, protein gene product 9.5; WAS, water avoidance stress.

Article Snippet: The sections were incubated with rabbit anti-rat polyclonal antibody raised against CHT1 (Millipore, Darmstadt, Germany) at room temperature for 24 hours, followed by incubation with the appropriate secondary antibodies.

Techniques: Immunofluorescence

Journal: PLoS ONE

Article Title: A Non-Neuronal Cardiac Cholinergic System Plays a Protective Role in Myocardium Salvage during Ischemic Insults

doi: 10.1371/journal.pone.0050761

Figure Lengend Snippet: (A) Compared with the right hindlimb (R), the left hindlimb subjected to IR had increased protein expression of ChAT and CHT1 in tissue samples isolated from quadriceps femoris muscle within 16 h ( P <0.05, n = 6, respectively). (B) The heart excised after IR also showed increased expression of both ChAT and CHT1, and the levels were peaked at 12 h (0 h vs. 12 h: P <0.05, n = 6, respectively) in the heart.

Article Snippet: The following primary antibodies were used: a goat anti-ChAT polyclonal antibody (Millipore) at 1∶500 dilution; a rabbit anti-CHT1 polyclonal antibody (Antagene, Inc., Limonest, France) at 1∶500 dilution; and a rabbit anti-cleaved caspase-3 monoclonal antibody (Cell Signaling Technology, Inc.) at 1∶500 dilution.

Techniques: Expressing, Isolation

Journal:

Article Title: Reduction in CHT1-mediated choline uptake in primary neurons from presenilin-1 M146V mutant knock-in mice

doi: 10.1016/j.brainres.2006.12.005

Figure Lengend Snippet: Table 1

Article Snippet: For immunocytochemical analysis of colocalization of CHT1 and choline acetyltransferase (ChAT) in primary neurons, the cultured cells were exposed to 1:1000 dilution of rabbit anti-CHT1 polyclonal antibody and 1:250 dilution of mouse anti-ChAT monoclonal antibody (Chemicon) overnight at 4 °C, followed by incubation for 1 h with a mixture of Texas Red-labeled anti-mouse and fluorescein-labeled anti-rabbit secondary antibodies (Vector Laboratories, Burlingame, California).

Techniques: Mutagenesis, Functional Assay

Journal: The Journal of Neuroscience

Article Title: Choline Transporter 1 Maintains Cholinergic Function in Choline Acetyltransferase Haploinsufficiency

doi: 10.1523/JNEUROSCI.1106-04.2004

Figure Lengend Snippet: Increased ChT activity, mRNA, and protein in Chat heterozygotes. a, [3H]Choline uptake was measured in the presence and absence of HC-3, an inhibitor of ChT activity, as described in Materials and Methods. The amount of ChT-transported choline was calculated by subtracting the amount of intracellular [3H]choline in the presence of HC-3 from the amount of [3H]choline in the absence of HC-3. The [3H]choline constituted a higher fraction of the HC-3-sensitive choline level in the Chat+/- mice compared with the wild-type mice (p < 0.05). ACh synthesized from [3H]choline was also measured in the presence and absence of HC-3. The amount of ACh synthesized from ChT-transported choline was calculated by subtracting the amount of ACh synthesized in the presence of HC-3 from the amount of ACh produced in the absence of HC-3. The newly made ACh constituted a higher fraction of the HC-3-sensitive ACh pool in the Chat+/- mice than it did in the wild-type littermates (p < 0.05). b, RT-PCR of CHT1 mRNA in Chat+/+ and Chat+/- mice. Septal RNA was used for RT-PCR of CHT1 and GAPDH. As expected, the PCR product from the amplification of CHT1 was present as an 840 bp band. The PCR product from the amplification of GAPDH was present as a 983 bp band. c, CHT1 product levels were quantified by Kodak Image Station Software and normalized to GAPDH product levels. The results are presented as a percentage of wild-type control ± range. d, CHT1 protein levels in Chat+/+ and Chat+/- mice. CHT1 protein levels in several CNS structures were measured by Western blot analysis. Chat+/- mice have an approximately twofold increase in CHT1 protein levels in each of the brain regions compared with Chat+/+ mice. e, CHT1 levels were quantified by Kodak Image Station Software and are presented as a percentage of wild-type control ± range. STR, Striatum; CTX, cerebral cortex; HPC, hippocampus; SC, spinal cord.

Article Snippet: After transfer of protein to an Immoblin P membrane (Millipore), the membrane was blocked with 5% nonfat dry milk in 1× Tris-buffered saline (TBS) containing 0.1% Tween 20 for 1 hr and then probed with affinity-purified rabbit anti-CHT1 polyclonal antibody AB5966 (1:1000; Chemicon, Temecula, CA) overnight.

Techniques: Activity Assay, Synthesized, Produced, Reverse Transcription Polymerase Chain Reaction, Amplification, Software, Western Blot